Screening of potential aquatic probiotics from the major microflora of guppies (Poecilia reticulata)

Aparna BALAKRISHNA, T. R. KEERTHI

PDF(306 KB)
PDF(306 KB)
Front. Chem. Sci. Eng. ›› 2012, Vol. 6 ›› Issue (2) : 163-173. DOI: 10.1007/s11705-012-1283-4
RESEARCH ARTICLE

Screening of potential aquatic probiotics from the major microflora of guppies (Poecilia reticulata)

Author information +
History +

Abstract

The fish (Poecilia reticulata) was used as the source for probiotics. 46 bacterial isolates were obtained from the skin, gills, guts and intestines of the guppy, Poecilia reticulata (collected from a government model fish farm in Kottayam, India). Of the above isolated strains, four isolates were selected based on their inhibitory spectrum against five indicator strains, Aeromonas hydrophila 1739, Vibrio cholerae 3906, Flavobacterium 2495, Acinetobacter 1271 and Alcaligenes 1424 (standard cultures collected from Microbial Type Culture Collection (MTCC) Chandigarh, India). Among the resulting isolates, two were gram-positive cocci, namely MBTU-PB2 and MBTU-PB3 and belong to the genus Staphylococcus. The other two were gram-negative rods, namely MBTU-PB1 and MBTU-PB4, of the genera Enterobacter and Acinetobacter, respectively. The basic probiotic characteristics of these isolates such as the production of bacteriocin like inhibitory substances (BLIS), antibiotic sensitivities and growth profiles were also determined. The above four isolated strains exhibited different antagonisms than the five indicator strains. During incubation, the antibacterial activity gradually increased in the inhibition zone and was influenced by the lag period (λ) and doubling time. The lag periods for most of the four selected strains were shorter than those of the indicator strains and the isolates had different growth rates (µ) than the indicator strains. All four isolates produced BLIS, however, the strains had different BLIS activities against the indicator strains. Treatment of the neutralized cell free supernatants of the selected isolates with proteases eliminated or reduced the BLIS activity, suggesting a proteinaceous nature of the inhibitory compounds. Further, the optimum BLIS activity was observed at neutral pH after 18 h of incubation. The antibiotic sensitivity assay revealed that the isolates were susceptible to routinely used antibiotics, whereas the plasmid profiles showed that the plasmids had no role in the antagonistic properties of the four isolated strains. The results showed that the isolates could be a promising source for biocontrol agents in aquacultures.

Keywords

aquaculture / normal flora / immunomodulatory effect / probiotic / antagonistic principle / bacteriocin like inhibitory substances

Cite this article

Download citation ▾
Aparna BALAKRISHNA, T. R. KEERTHI. Screening of potential aquatic probiotics from the major microflora of guppies (Poecilia reticulata). Front Chem Sci Eng, 2012, 6(2): 163‒173 https://doi.org/10.1007/s11705-012-1283-4

References

[1]
Maeda M, Liao I C. Microbial processes in aquaculture environment and their importance for increasing crustacean production. Japan Agricultural Research Quarterly, 1994, 28: 283–288
[2]
Cannell R J P, Owsianka A M, Walker J M. Results of a large scale screening programme to detect antibacterial activity from fresh water algae. British Phycological Journal, 1988, 23(1): 41–44
CrossRef Google scholar
[3]
Grisez L, Ollevier F. Vibrio (Listonella) Anguillarum Infection in Marine Fish Larviculture. In: Lavens P, Jaspers E, Roelands I, eds. Larvi’91–Fish and Crustacean Larviculture Symposium. European Aquaculture Society, Gent, 1995, 24: 497
[4]
Hansen G H, Olafsen J A. Bacterial interactions in early life stages of marine cold water fish. Microbial Ecology, 1999, 38(1): 1–26
CrossRef Pubmed Google scholar
[5]
Verschuere L, Rombaut G, Sorgeloos P, Verstraete W. Probiotic bacteria as biological control agents in aquaculture. Microbiology and Molecular Biology Reviews, 2000, 64(4): 655–671
CrossRef Pubmed Google scholar
[6]
Schwarz S, Kehrenberg C, Walsh T R. Use of antimicrobial agents in veterinary medicine and food animal production. International Journal of Antimicrobial Agents, 2001, 17(6): 431–437
CrossRef Pubmed Google scholar
[7]
Akinbowale O L, Peng H, Barton M D. Antimicrobial resistance in bacteria isolated from aquaculture sources in Australia. Journal of Applied Microbiology, 2006, 100(5): 1103–1113
CrossRef Pubmed Google scholar
[8]
Moriarty D J W. Control of luminous Vibrio species in penaeid aquaculture ponds. Aquaculture (Amsterdam, Netherlands), 1998, 164(1–4): 351–358
CrossRef Google scholar
[9]
Vine N G, Leukes W D, Kaiser H. In vitro growth characteristics of five candidate aquaculture probiotics and two fish pathogens grown in fish intestinal mucus. FEMS Microbiology Letters, 2004, 231(1): 145–152
CrossRef Pubmed Google scholar
[10]
Fuller R. History and Development of Probiotics. In: Fuller R, ed. Probiotics: The Scientific Basis. London: Chapman and Hall, 1992, 1–8
[11]
Austin B, Stuckey L F, Robertson P A W, Effendi I, Griffith D R W. A probiotic strain of Vibrio alginolyticus effective in reducing diseases caused by Aeromonas salmonicida, Vibrio anguillarum and Vibrio ordalii. Journal of Fish Diseases, 1995, 18(1): 93–96
CrossRef Google scholar
[12]
Moriarty D J W. Diseases Control in Shrimp Aquaculture with Probiotic Bacteria. In: Bell C R, Brylinsky M, Johnson-Green P, eds. Microbial Biosystems: New Frontiers. Proceedings of the 8th International Symposium on Microbial Ecology. Atlantic Canada Society for Microbial Ecology, Halifax, Canada, 1999, 237–243
[13]
Atlas R M, Bartha E. Microbial Ecology Fundamentals and Application. Menlo Park: Benjamin Cummings Science Publishing, 1997
[14]
Gould G. Industry perspectives on the use of natural antimicrobials and inhibitors for food applications. Journal of Food Protection, 1996, 59: S82–S86
[15]
McAuliffe O, Ross R P, Hill C. Lantibiotics: structure, biosynthesis and mode of action. FEMS Microbiology Reviews, 2001, 25(3): 285–308
CrossRef Pubmed Google scholar
[16]
Tagg J R, Dajani A S, Wannamaker L W. Bacteriocins of gram-positive bacteria. Bacteriological Reviews, 1976, 40(3): 722–756
Pubmed
[17]
Riley M A, Wertz J E. Bacteriocins: evolution, ecology, and application. Annual Review of Microbiology, 2002, 56(1): 117–137
CrossRef Pubmed Google scholar
[18]
Dopazo C P, Lemos M L, Lodeiros C, Bolinches J, Barja J L, Toranzo A E. Inhibitory activity of antibiotic-producing marine bacteria against fish pathogens. Journal of Applied Bacteriology, 1988, 65(2): 97–101
CrossRef Pubmed Google scholar
[19]
Anne J A, Praseeja R J, Keerthi T R. Evaluation of probiotic potential of lactic acid bacteria isolated from infant feces and the study of its effect on the enteric fever pathogens— Salmonella typhi and Salmonella paratyphi A. International Journal of Chemical Science, 2010, 8(5): S376– S387
[20]
Maniatis T, Fritsch E F, Sambrook J. Molecular Cloning. A Laboratory Manual. New York: Cold Spring Harbour Laboratory, Cold Spring Harbour, 1982
[21]
Trevors J T. Plasmid curing in bacteria. FEMS Microbiology Letters, 1986, 32(3–4): 149–157
CrossRef Google scholar
[22]
Zar J H. Biostatical Analysis. New Jersey: Prentice-Hall, Englewood Cliffs, 1984, 2: 718
[23]
Zwietering M H, Jongenburger I, Rombouts F M, van’t Riet K. Modeling of the bacterial growth curve. Applied and Environmental Microbiology, 1990, 56(6): 1875–1881
Pubmed
[24]
Sugita H, Matsuo N, Shibuya K, Degunchi Y. Production of antibacterial substances by intestinal bacteria isolated from coastal crab and fish species. Journal of Marine Biotechnology, 1996, 4: 220–223
[25]
Azad I S, Al-Marzouk A. Autochthonous aquaculture probiotics—a critical analysis. Research Journal of BioTechnology, 2008, 3: 171–177
[26]
Lazado C C, Caipang C M A, Rajan B, Brinchmann M F, Kiron V. Characterization of GP21 and GP12: two potential probiotic bacteria isolated from the gastrointestinal tract of Atlantic Cod. Probiotics & Antimicrobial Proteins, 2010, 2(2): 126–134
CrossRef Google scholar
[27]
Balcazar J L, de Blas I, Ruiz-Zarzuela I, Vendrell D, Muzquiz J L. Probiotics: a tool for the future of fish and shellfish health management. Journal of Aquaculture in the Tropics, 2004, 19: 239–242
[28]
Geovanny D G R, Balcázar J L, Ma S. Probiotics as control agents in aquaculture. Journal of Ocean University of China, 2007, 6(1): 76–79 (in Chinese)
CrossRef Google scholar
[29]
de Vuyst L, Vandamme E J. Bacteriocins of Lactic Acid Bacteria. London: Blackie Academic & Professional, 1994, 91–142
[30]
Salinas I, Cuesta A, Esteban M A, Meseguer J. Dietary administration of actobacillus delbrüeckiiL and Bacillus subtilis, single or combined, on gilthead seabream cellular innate immune responses. Fish & Shellfish Immunology, 2005, 19(1): 67–77
CrossRef Pubmed Google scholar
[31]
Parente E, Ricciardi A. Production, recovery and purification of bacteriocins from lactic acid bacteria. Applied Microbiology and Biotechnology, 1999, 52(5): 628–638
CrossRef Pubmed Google scholar
[32]
Duitman E H, Hamoen L W, Rembold M, Venema G, Seitz H, Saenger W, Bernhard F, Reinhardt R, Schmidt M, Ullrich C, Stein T, Leenders F, Vater J. The mycosubtilin synthetase of Bacillus subtilis ATCC6633: a multifunctional hybrid between a peptide synthetase, an amino transferase, and a fatty acid synthase. Proceedings of the National Academy of Sciences of the United States of America, 1999, 96(23): 13294–13299
CrossRef Pubmed Google scholar
[33]
Bizani D, Brandelli A. Characterization of a bacteriocin produced by a newly isolated Bacillus sp. Strain 8 A. Journal of Applied Microbiology, 2002, 93(3): 512–519
CrossRef Pubmed Google scholar
[34]
Ghanbari M, Rezaei M, Soltani M, Shah-Hosseini G. Production of bacteriocin by a novel Bacillus sp. strain RF 140, an intestinal bacterium of Caspian Frisian Roach (Rutilus frisii kutum). Iranian Journal of Veterinary Research, 2009, 10(3): 267–272
[35]
Pinchuk I V, Bressollier P, Verneuil B, Fenet B, Sorokulova I B, Mégraud F, Urdaci M C. In vitro anti-Helicobacter pylori activity of the probiotic strain Bacillus subtilis 3 is due to secretion of antibiotics. Antimicrobial Agents and Chemotherapy, 2001, 45(11): 3156–3161
CrossRef Pubmed Google scholar
[36]
Messi P, Guerrieri E, Bondi M. Bacteriocin-like substance (BLS) production in Aeromonas hydrophila water isolates. FEMS Microbiology Letters, 2003, 220(1): 121–125
CrossRef Pubmed Google scholar
[37]
Lee K H, Jun K D, Kim W S, Paik H D. Partial characterization of polyfermenticin SCD, a newly identified bacteriocin of Bacillus polyfermenticus. Letters in Applied Microbiology, 2001, 32(3): 146–151
CrossRef Pubmed Google scholar
[38]
Patel A K, Deshattiwar M K, Chaudhari B L, Chincholkar S B. Production, purification and chemical characterization of the catecholate siderophore from potent probiotic strains of Bacillus spp. Bioresource Technology, 2009, 100(1): 368–373
CrossRef Pubmed Google scholar

Acknowledgments

The authors gratefully acknowledge financial support from the University Grants Commission, Government of India, New Delhi, under Grant UGC-F.No.37-264/2009 (SR).

RIGHTS & PERMISSIONS

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg
AI Summary AI Mindmap
PDF(306 KB)

Accesses

Citations

Detail

Sections
Recommended

/